Yeast cell expressing heterologous receptor kinase

Abstract

Disclosed herein are compositions and methods which are useful in the identification and isolation of components involved in transmembrane receptor-mediated signaling. Such components include the receptors themselves (e.g., tyrosine kinase receptors, cytokine receptors and tyrosine phosphatase receptors), as well as ligands which bind the receptors and modulators of the downstream intracellular catalytic event which characterizes receptor-mediated signalling.

We claim: 
     
       1. A yeast cell containing an expressible copy of a gene encoding a heterologous transmembrane receptor, the intracellular domain of the heterologous transmembrane receptor exhibiting tyrosine kinase activity in response to ligand binding. 
     
     
       2. A yeast cell of claim 1 which is transformed with a CEN-based plasmid which contains the expressible copy of the gene encoding the heterologous transmembrane receptor. 
     
     
       3. A yeast cell of claim 1 which contains a chromosome which has been modified by the insertion of the gene encoding the heterologous transmembrane receptor. 
     
     
       4. A yeast cell of claim 1 wherein the heterologous transmembrane receptor gene is derived from a mammalian source. 
     
     
       5. A yeast cell of claim 1 which is transformed with a cDNA library constructed from an organism of interest. 
     
     
       6. A yeast cell of claim 5 wherein the cDNA library is under the control of an inducible promoter. 
     
     
       7. A yeast cell of claim 1 wherein the gene encoding the heterologous transmembrane receptor is carried on a CEN-based plasmid. 
     
     
       8. A CEN-based plasmid carrying a gene encoding a heterologous transmembrane receptor, the heterologous transmembrane receptor, when expressed in yeast, being characterized by the ability to exhibit tyrosine kinase activity in response to ligand binding. 
     
     
       9. A yeast cell containing a heterologous transmembrane receptor, the heterologous transmembrane receptor exhibiting tyrosine kinase activity in response to ligand binding. 
     
     
       10. A yeast cell of claim 9 which is transformed with a cDNA library constructed from an organism of interest. 
     
     
       11. A yeast cell of claim 10 wherein the cDNA library is under the control of an inducible promoter. 
     
     
       12. A yeast cell containing an expressible copy of a gene encoding a heterologous transmembrane receptor, the intracellular domain of the heterologous transmembrane receptor being characterized by the ability to activate a heterologous cytoplasmic tyrosine kinase activity in response to ligand binding. 
     
     
       13. A yeast cell of claim 13 wherein the heterologous transmembrane receptor is a cytokine receptor. 
     
     
       14. A yeast cell of claim 13 wherein the gene encoding the heterologous transmembrane receptor is carried on a CEN-based plasmid. 
     
     
       15. A yeast cell of claim 13 which contains a chromosome which has been modified by the insertion of the gene encoding the heterologous transmembrane receptor. 
     
     
       16. A yeast cell of claim 13 wherein the heterologous transmembrane receptor gene is derived from a mammalian source. 
     
     
       17. A yeast cell of claim 13 which is transformed with a cDNA library constructed from an organism of interest. 
     
     
       18. The yeast cell of claim 17 wherein the cDNA library is under the control of an inducible promoter. 
     
     
       19. A CEN-based plasmid carrying a gene encoding a heterologous transmembrane receptor, the heterologous transmembrane receptor being characterized by the ability to activate a cytoplasmic tyrosine kinase activity in response to ligand binding. 
     
     
       20. A yeast cell containing a heterologous transmembrane receptor, the heterologous transmembrane receptor being characterized by the ability to activate a heterologous cytoplasmic tyrosine kinase activity in response to ligand binding. 
     
     
       21. A yeast cell of claim 20 wherein the heterologous transmembrane receptor is a cytokine receptor. 
     
     
       22. A yeast cell of claim 20 which is transformed with a cDNA library constructed from an organism of interest. 
     
     
       23. The yeast cell of claim 22 wherein the cDNA library is under the control of an inducible promoter. 
     
     
       24. A yeast cell containing an expressible copy of a gene encoding a chimeric heterologous transmembrane receptor comprising the intracellular domain of a tyrosine kinase receptor fused to the extracellular domain of a second transmembrane receptor, the intracellular domain of the chimeric heterologous transmembrane receptor exhibiting tyrosine kinase activity in response to ligand binding. 
     
     
       25. A yeast cell of claim 24 wherein the extracellular domain is from a cytokine receptor. 
     
     
       26. A yeast cell of claim 24 wherein the extracellular domain is from a tyrosine phosphatase receptor. 
     
     
       27. A yeast cell of claim 24 which is transformed with a cDNA library constructed from an organism of interest. 
     
     
       28. The yeast cell of claim 27 wherein the cDNA library is under the control of an inducible promoter. 
     
     
       29. A yeast cell containing an expressible copy of a gene encoding a chimeric heterologous transmembrane receptor comprising an intracellular domain fused to the extracellular domain of a second transmembrane receptor, the intracellular domain of the chimeric heterologous transmembrane receptor being characterized by the ability to activate a heterologous cytoplasmic tyrosine kinase activity in response to ligand binding. 
     
     
       30. A yeast cell of claim 29 wherein the extracellular domain is from a cytokine receptor. 
     
     
       31. A yeast cell of claim 29 wherein the extracellular domain is from a tyrosine phosphatase receptor. 
     
     
       32. A yeast cell of claim 29 which is transformed with a cDNA library constructed from an organism of interest. 
     
     
       33. A yeast cell of claim 32 wherein the cDNA library is under the control of an inducible promoter. 
     
     
       34. A yeast cell containing an expressible copy of a gene encoding a heterologous transmembrane receptor selected from the group consisting of a tyrosine kinase receptor, and a tyrosine phosphatase receptor.